Our continuing and principal aim is the elucidation of the physical chemistry of the serum lipoproteins in states of health and disease. To achieve this objective, we anticipate defining in greater detail the physical characteristics and chemical composition of each serum lipoprotein class as improvements in techniques for lipoprotein isolated and analysis permit. Such studies, in addition, should provide insight into the factors that determine lipoprotein composition and fluctuations in composition within each lipoprotein class. Substances to be analyzed include protein, the major classes of lipids, their associated fatty acids, as well as hydrocarbons, steroids, and other substances which may be present as part of the lipoprotein structure. Particular emphasis will be given the precise quantitative analysis of lipoprotein protein and its relationship to lipoprotein physical and chemical properties within all subfractions. Special efforts will be made to develop and refine ultracentrifugal techniques for the isolation and subfractionation of all classes of serum lipoproteins in as uncontaminated and native form as is possible. "Cumulative flotation rate" techniques, on both dynamic and equilibrium density gradients, will be an important method employed. Precise physical and chemical analysis of such subfractions will provide important insights into lipoprotein structure within each lipoprotein class. Also, methods both simple and complex will be developed and improved for reliable serum lipid and lipoprotein analysis. We are particularly concerned with the refinement and utilization of a simple, inexpensive, quantitative, agarose gel lipoprotein electrophoresis test. Application of our developed techniques will involve active biomedical collaboration with other scientists, especially those having access to unusual clinical resources and to special animal populations. All studies will use computer techniques and appropriate instrumentation.